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Abstract：
Artificial insemination in queen honey bees is the only tool that provides complete control over mating for research and breeding purposes, making it essential in genetic improvement and conservation programs in this species. The aims of this study were to characterize drone semen bacterial loads by culture-dependent and independent methods and to describe their variation depending on the method of semen collection, the colony and the apiary. In the first experiment, the bacterial loads of semen collected from the seminal vesicles or from ejaculates was studied using culture-dependent methods. The collection method had a significant influence on the overall bacterial count in semen. Out of the 42 semen samples analyzed, 26 (61.9%) tested positive for bacterial isolation. This encompassed the entirety of samples obtained from the seminal vesicles (21 of 21), whereas only 23.8% of those derived from ejaculates (5 out of 21) showed bacterial isolation. In the second experiment, next-generation sequencing techniques were used to describe the microbiome of ejaculated drone semen for the first time. The most abundant phyla were Proteobacteria, Firmicutes, Bacteroidota and Actinobacteriota, while the most abundant genera were Lactobacillus, Staphylococcus, Prevotella, Alloprevotella and Streptococcus. The results showed that the apiary had a significant effect on the community structure composition and abundance of the seminal microbiota, and significative differences in abundance were observed for the genera Sphingomonas, Methylobacterium-Methylorubrum, Bifidobacterium and Alloprevotella. Significant differences were also observed in the richness of the microbiota between apiaries and colonies.
摘要：
蜂王蜜蜂的人工授精是为研究和繁殖目的提供完全控制交配的唯一工具，使其在该物种的遗传改良和保护计划中必不可少。这项研究的目的是通过培养依赖和独立的方法来表征无人机精液细菌负荷，并描述它们根据精液收集方法的变化。殖民地和养蜂场。在第一个实验中,使用培养依赖性方法研究了从精囊或射精中收集的精液的细菌负荷。收集方法对精液中的总体细菌计数有显著影响。在分析的42份精液样本中，26例（61.9%）细菌分离检测呈阳性。这包括从精囊获得的全部样品（21个中的21个），而来自射精的只有23.8％（21个中的5个）显示细菌分离。在第二个实验中,新一代测序技术首次用于描述射精无人机精液的微生物组.最丰富的门是变形杆菌，Firmicutes,类细菌和放线菌,虽然最丰富的属是乳酸菌，葡萄球菌,普雷沃氏菌,异普氏菌和链球菌。结果表明:养蜂场对群落结构组成和精液微生物的丰度有显著影响,并且观察到鞘氨醇单胞菌属的丰度存在显着差异，甲基细菌-甲基细菌，双歧杆菌和Alloprevotella。还观察到在蜂巢和菌落之间的微生物群的丰富度存在显着差异。
